Rapid Sample Preparation and Ultrasensitive Detection of COVID-19 in Human Saliva

人类唾液中 COVID-19 的快速样品制备和超灵敏检测

• 批准号: 10286212
• 负责人: Tak Sing Wong
• 金额: $ 22.92万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10286212
• 关键词: 2019-nCoV; Antibodies; Biological Markers; Biological Models; COVID-19; COVID-19 detection; COVID-19 patient; COVID-19 screening; COVID-19 test; Cessation of life; Coffee; DNA; Detection; Diagnostic tests; Disease Outbreaks; Engineering; Enzyme-Linked Immunosorbent Assay; Goals; Hour; Human; Human Resources; Libraries; Measures; Methods; Molecular Target; Nature; Patients; Performance; Polymerase Chain Reaction; Preparation; RNA; Rapid screening; Reagent; Research; Reverse Transcription; SARS-CoV-2 infection; Saliva; Sampling; Sensitivity and Specificity; Speed; Surface; Testing; Time; Training; United States; Viral; Viral Antibodies; Virion; Virus; antibody test; antigen test; aptamer; convolutional neural network; cost; deep learning; design; detection limit; detection platform; disorder control; multiplex detection; nanoparticle; neural network algorithm; novel; pandemic disease; particle; targeted biomarker; viral RNA; viral detection

Project Summary The coronavirus disease 2019 (COVID-19) – caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) – is a global pandemic with worldwide cases of over 35 million and a death toll of over 1 million people. In the United States, there are a total of over 7.5 million cases with over 210,000 deaths as of October 2020. The ability to identify COVID-19 infected patients rapidly, accurately, and cost-effectively is of paramount importance to control the disease outbreak. The two different types of COVID-19 tests are diagnostic tests and antibody tests. Specifically, the diagnostic tests can only identify active COVID-19 infected patients; the antibody tests can only indicate prior COVID-19 infections but not active cases. These diagnostic tests include quantitative reverse transcription polymerase chain reaction (qRT-PCR) and rapid antigen tests. In particular, the qRT-PCR is currently the most sensitive method (down to ~100 copies/mL) but can take >12 hours to perform and require trained personnel, specific reagents, and expensive machines. The rapid antigen tests provide results within 30 minutes, but these tests require the samples to contain high virus concentration (i.e., >107 virus particles/mL) leading to increasing chances of false-negative cases. In addition, one common example of the antibody tests is the enzyme-linked immunosorbent assay (ELISA). This test only applies to patients who have already developed the antibodies, which could take several days to weeks. Thus far, no single tests can detect both active and previously COVID-19 infected cases. The objective of the proposed research is to develop a proof-of-concept rapid and ultrasensitive virus-detection platform termed viral-ID, which is capable of simultaneous detecting the SARS-CoV-2 RNA and antibodies inside saliva for the identification of both active and previous COVID-19 infections within a single test. Specifically, the viral-ID platform will allow for specific molecular targeting, separation, enrichment, and filtering, all carried out within a single saliva droplet utilizing the nature-inspired coffee ring effect. Targeting and detection of the viral biomarkers will be carried out by aptamer-functionalized nanoparticles and deep-learning-enabled surface- enhanced Raman scattering (SERS), respectively. Using synthetic SARS-CoV-2 RNA and antibody-spiked saliva as a model system, the goal of this research is to demonstrate a bio-analysis platform capable of rapid sample preparation and ultrasensitive detection of both viral RNA (~10 – 100 copies/mL) and antibodies (~femtomolar) in a saliva droplet within 30 minutes. If successful, our viral-ID platform will provide all-in-one and best-in-class diagnostic and antibody test in terms of speed, sensitivity, and specificity for the rapid screening of COVID-19.

项目摘要 冠状病毒病2019(新冠肺炎)-由新型严重急性呼吸系统综合症引起 冠状病毒2(SARS-CoV-2)-是一种全球性大流行，全球病例超过3500万例，死亡人数 超过100万人。在美国，总共有超过750万例病例，死亡人数超过21万人 截至2020年10月。 快速、准确和经济高效地识别新冠肺炎感染患者的能力至关重要 控制疾病暴发的重要性。新冠肺炎测试的两种不同类型是诊断性测试和 抗体测试。具体地说，诊断测试只能识别活跃的新冠肺炎感染患者；抗体 检测只能显示以前感染新冠肺炎的病例，而不能显示活跃的病例。这些诊断测试包括定量 逆转录-聚合酶链式反应(qRT-PCR)和快速抗原检测。特别是，qRT-PCR 是目前最敏感的方法(低至~100份/毫升)，但可能需要&gt；12小时才能完成并需要 训练有素的人员、特殊的试剂和昂贵的机器。快速抗原测试在30分钟内得出结果 几分钟，但这些测试要求样本含有较高的病毒浓度(即&gt；107病毒颗粒/毫升) 导致假阴性病例的机会增加。此外，抗体测试的一个常见例子 是酶联免疫吸附试验(EL ISA)。这项测试只适用于已经 研制出了抗体，这可能需要几天到几周的时间。到目前为止，没有一项测试可以同时检测到这两种情况 活跃的和以前感染过新冠肺炎的病例。 这项研究的目标是开发一种概念验证、快速和超灵敏的病毒检测方法。 名为Virus-ID的平台，能够同时检测SARS-CoV-2RNA和内部抗体 唾液用于在一次测试中识别活跃的和既往的新冠肺炎感染。具体地说， 病毒识别平台将允许特定的分子靶向、分离、浓缩和过滤，所有这些都可以进行 利用受自然启发的咖啡戒指效应，只需一滴唾液即可。病毒的靶向和检测 生物标记物将通过适体功能化的纳米颗粒和支持深度学习的表面- 增强拉曼散射(SERS)。使用合成的SARS-CoV-2 RNA和抗体添加 以唾液为模型系统，本研究的目标是展示一种能够快速 病毒RNA(~10-100拷贝/毫升)和抗体的样品制备及超灵敏检测 (~飞摩尔)在30分钟内在唾液滴中。如果成功，我们的病毒ID平台将提供一体机和 在速度、灵敏度和特异性方面是同类中最好的诊断和抗体测试，用于快速筛查 新冠肺炎。